Table 3.
PCR primers used for site-directed mutagenesis.
| Primer sequence | Selection | Mutant |
| 5'-ACCGGCCTGGGCCGGCCCG(CA)GATGGCGGTCTACCAC-3' |
BstUI (mutant D58A) |
D58A D58E |
| 5'-GGTGGTAGACCGCCATC(TG)CGGGCCGGCCCAGGCCGG-3' | ||
| 5'-TCCTGGTGGGCTTCGTG(ACGT)(ACGT)(ACGT)CTCAAGGCTCCGGGCAAG-3' |
SacI restriction site disappears |
E72X |
| 5'-CTTGCCCGGAGCCTTGAG(ACGT)(ACGT)(ACGT)CACGAAGCCCACCAGGA-3' | ||
| 5'-CTGGTGGGCTTCGTGGAGCTC(GC)(GC)GGCCCCGGGCAGG-3' |
SmaI/ApaI (K74G, K74R) |
K74G K74A K74R K74P |
| 5'-CTTGCCCGGGGCC(GC)(GC)GAGCTCCACGAAGCCCACAGG-3' | ||
| 5'-GTTACCGTGATAGACCCGGCCATGGGCACGGGGACCTT-3' |
NcoI (V356M) |
V356M |
| 5'-AAGGTCCCCGTGCCCATGGCCGGGTCTATCACGGTAAC-3' | ||
| 5'-TGGTGATCCTTGGCGCCCCCCCCTACGACCGGGTG-3' |
NarI (N473A) |
N473A |
| 5'-CACCCGGTCGTAGGGGGGGGCGCCAAGGATCACCA-3' | ||